RESUMO
A major question in evolutionary biology is how mating patterns affect the fitness of offspring. However, in animals and seed plants it is virtually impossible to investigate the effects of specific gamete genotypes. In bryophytes, haploid gametophytes grow via clonal propagation and produce millions of genetically identical gametes throughout a population. The main goal of this research was to test whether gamete identity has an effect on the fitness of their diploid offspring in a population of the aquatic peat moss Sphagnum macrophyllum. We observed a heavily male-biased sex ratio in gametophyte plants (ramets) and in multilocus microsatellite genotypes (genets). There was a steeper relationship between mating success (number of different haploid mates) and fecundity (number of diploid offspring) for male genets compared with female genets. At the sporophyte level, we observed a weak effect of inbreeding on offspring fitness, but no effect of brood size (number of sporophytes per maternal ramet). Instead, the identities of the haploid male and haploid female parents were significant contributors to variance in fitness of sporophyte offspring in the population. Our results suggest that intrasexual gametophyte/gamete competition may play a role in determining mating success in this population.
Assuntos
Diploide , Aptidão Genética , Haploidia , Sphagnopsida/genética , Fertilidade , Variação Genética , Genótipo , Células Germinativas Vegetais/fisiologia , Endogamia , Repetições de Microssatélites , South Carolina , Sphagnopsida/fisiologiaRESUMO
Morphological characters from the gametophyte and sporophyte generations have been used in land plants to infer relationships and construct classifications, but sporophytes provide the vast majority of data for the systematics of vascular plants. In bryophytes both generations are well developed and characters from both are commonly used to classify these organisms. However, because morphological traits of gametophytes and sporophytes can have different genetic bases and experience different selective pressures, taxonomic emphasis on one generation or the other may yield incongruent classifications. The moss order Hookeriales has a controversial taxonomic history because previous classifications have focused almost exclusively on either gametophytes or sporophytes. The Hookeriales provide a model for comparing morphological evolution in gametophytes and sporophytes, and its impact on alternative classification systems. In this study we reconstruct relationships among mosses that are or have been included in the Hookeriales based on sequences from five gene regions, and reconstruct morphological evolution of six sporophyte and gametophyte traits that have been used to differentiate families and genera. We found that the Hookeriales, as currently circumscribed, are monophyletic and that both sporophyte and gametophyte characters are labile. We documented parallel changes and reversals in traits from both generations. This study addresses the general issue of morphological reversals to ancestral states, and resolves novel relationships in the Hookeriales.
Assuntos
Bryopsida/classificação , Bryopsida/genética , Células Germinativas Vegetais/fisiologia , Filogenia , Sequência de Bases , Evolução Biológica , Bryopsida/anatomia & histologia , DNA de Plantas/genética , DNA Espaçador Ribossômico/genética , Evolução Molecular , Células Germinativas Vegetais/classificação , Mitocôndrias/genética , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Polyploidization is thought to result in instant sympatric speciation, but several cases of hybrid zones between one of the parental species and its polyploid derivative have been documented. Previous work showed that diploid Sphagnum lescurii is an allopolyploid derived from the haploids S. lescurii (maternal progenitor) and S. subsecundum (paternal progenitor). Here, we report the results from analyses of a population where allodiploid and haploid S. lescurii co-occur and produce sporophytes. We tested (i) whether haploids and diploids form hybrid triploid sporophytes; (ii) how hybrid and nonhybrid sporophytes compare in fitness; (iii) whether hybrid sporophytes form viable spores; (iv) the ploidy of any viable gametophyte offspring from hybrid sporophytes; (v) the relative viability of sporelings derived from hybrid and nonhybrid sporophytes; and (vi) if interploidal hybridization results in introgression between the allopolyploid and its haploid progenitor. We found that triploid hybrid sporophytes do occur and are larger than nonhybrid sporophytes, but exhibit very low germination percentages and produce sporelings that develop more slowly than those from nonhybrid sporophytes. All sporophytes attached to haploid gametophytes were triploid and were sired by diploid males, but all sporophytes attached to diploid gametophytes were tetraploid. This asymmetric pattern of interploidal hybridization is related to an absence of haploid male gametophytes in the population. Surprisingly, all sporelings from triploid sporophytes were triploid, yet were genetically variable, suggesting some form of aberrant meiosis that warrants further study. There was limited (but some) evidence of introgression between allodiploid and haploid S. lescurii.
Assuntos
Células Germinativas Vegetais/fisiologia , Hibridização Genética , Isolamento Reprodutivo , Sphagnopsida/genética , DNA de Plantas/genética , Aptidão Genética , Genética Populacional , Genoma de Planta , Desequilíbrio de Ligação , Repetições de Microssatélites , Ploidias , Reprodução/genética , Sphagnopsida/fisiologiaRESUMO
The monoicous peatmoss Sphagnum subnitens has a tripartite distribution that includes disjunct population systems in Europe (including the Azores), northwestern North America and New Zealand. Regional genetic diversity was highest in European S. subnitens but in northwestern North America, a single microsatellite-based multilocus haploid genotype was detected across 16 sites ranging from Coos County, Oregon, to Kavalga Island in the Western Aleutians (a distance of some 4115 km). Two multilocus haploid genotypes were detected across 14 sites on South Island, New Zealand. The microsatellite-based regional genetic diversity detected in New Zealand and North American S. subnitens is the lowest reported for any Sphagnum. The low genetic diversity detected in both of these regions most likely resulted from a founder event associated with vegetative propagation and complete selfing, with one founding haploid plant in northwest North America and two in New Zealand. Thus, one plant appears to have contributed 100% of the gene pool for the population systems of S. subnitens occurring in northwest North America, and this is arguably the most genetically uniform group of plants having a widespread distribution yet detected. Although having a distribution spanning 12.5° of latitude and 56° of longitude, there was no evidence of any genetic diversification in S. subnitens in northwest North America. No genetic structure was detected among the three regions, and it appears that European plants of S. subnitens provided the source for New Zealand and northwest North American populations.
Assuntos
Pool Gênico , Genética Populacional , Haploidia , Sphagnopsida/genética , Alelos , DNA de Plantas/genética , Europa (Continente) , Efeito Fundador , Variação Genética , Genótipo , Desequilíbrio de Ligação , Repetições de Microssatélites , Nova Zelândia , América do Norte , Análise de Componente Principal , Análise de Sequência de DNARESUMO
It is commonly found that individual hybrid, polyploid species originate recurrently and that many polyploid species originated relatively recently. It has been previously hypothesized that the extremely rare allopolyploid peat moss Sphagnum troendelagicum has originated multiple times, possibly after the last glacial maximum in Scandinavia. This conclusion was based on low linkage disequilibrium in anonymous genetic markers within natural populations, in which sexual reproduction has never been observed. Here we employ microsatellite markers and chloroplast DNA (cpDNA)-encoded trnG sequence data to test hypotheses concerning the origin and evolution of this species. We find that S. tenellum is the maternal progenitor and S. balticum is the paternal progenitor of S. troendelagicum. Using various Bayesian approaches, we estimate that S. troendelagicum originated before the Holocene but not before c. 80,000 years ago (median expected time since speciation 40 000 years before present). The observed lack of complete linkage disequilibrium in the genome of this species suggests cryptic sexual reproduction and recombination. Several lines of evidence suggest multiple origins for S. troendelagicum, but a single origin is supported by approximate Bayesian computation analyses. We hypothesize that S. troendelagicum originated in a peat-dominated refugium before last glacial maximum, and subsequently immigrated to central Norway by means of spore flow during the last thousands of years.
Assuntos
Evolução Molecular , Sphagnopsida/genética , Teorema de Bayes , Cloroplastos/genética , Marcadores Genéticos , Especiação Genética , Genótipo , Haplótipos , Repetições de Microssatélites , Noruega , Filogeografia , Polimorfismo GenéticoRESUMO
Immature and mature B cells differ in the signals generated and transduced through their antigen receptor, surface immunoglobulin M (sIgM). Whereas signals generated through sIgM on mature B cells initiate a program leading to the positive activation of these cells, signaling through this receptor at the immature stage of development leads to a state of induced unresponsiveness or tolerance. Our previous studies have described developmental differences in sIgM transmembrane signaling that are independent of ligand-receptor affinity. In an attempt to understand the molecular basis for signaling differences between immature and mature B cells, we have analyzed the sIgM receptor complex in neonatal and adult mouse splenic B cells. While previously described components of this complex do not exhibit marked developmentally regulated differences in their association with sIgM, we have identified a 56-kD protein that associates with sIgM in mature (antigen-responsive), but not immature (tolerance-sensitive) B cells. This protein (p56) associates with sIgM as a homodimer, is constitutively phosphorylated on tyrosine, and is coimmunoprecipitated with IgM but not IgD. The observed inability to iodinate p56 suggests it is an intracellular component of the receptor complex. Based upon its migration in one- and two-dimensional gel electrophoresis we show, however, that p56 is distinct from the blk, lyn, or fyn src family kinases that have been shown to be associated with sIgM in mature B cells. The developmentally regulated participation of p56 in the B cell antigen receptor complex suggests a role in the differential signaling mediated via sIgM on immature and mature B cells.
Assuntos
Linfócitos B/imunologia , Imunoglobulina M/metabolismo , Receptores de Antígenos de Linfócitos B/metabolismo , Receptores Fc/análise , Animais , Imunoglobulina D/metabolismo , Camundongos , Peso Molecular , Testes de Precipitina , Proteínas Tirosina Quinases/análise , CoelhosRESUMO
The transporter associated with antigen presentation (TAP) complex shuttles cytosolic peptides into the exocytic compartment for association with nascent major histocompatibility complex class I molecules. Biochemical studies of murine and human TAP have established that substrate length and COOH-terminal residue identity are strong determinants of transport efficiency. However, the existence of these specificities in the intact cell and their influences on T cell responses have not been demonstrated. We have devised a method for studying TAP- mediated transport in intact cells, using T cell activation as a readout. The approach makes use of a panel of recombinant vaccinia viruses expressing peptides containing the Kd-restricted nonamer influenza nucleoprotein residues 147-155. The COOH terminus of each construct was appended with a dipeptide composed of an internal threonine residue followed by a varying amino acid. Synthetic peptide versions of these 11-mers exhibit vastly different transport capabilities in streptolysin O-permeabilized cells, in accordance with the predicted influence of the COOH-terminal residues. Presentation of the endogenously expressed version of each construct requires TAP-mediated transport and cooexpression with a vac-encoded exocytic COOH-terminal dipeptidase, angiotensin converting enzyme, to allow liberation of the minimal epitope. Recognition by epitope-specific CTLs therefore signifies TAP-mediated transport of a complete 11-mer within the target cell. Under normal assay conditions no influences of the COOH-terminal residue were revealed. However, when T cell recognition was limited, either by blocking CD8 coreceptor interactions or by decreasing the amount of transport substrate synthesized, significant COOH-terminal effects were revealed. Under such conditions, those peptides that transported poorly in biochemical assays were less efficiently presented. Therefore, TAP specificity operates in the intact cell, appears to reflect previously defined rules with regard to the influence of the COOH-terminal residue, and can strongly influence T cell responses.
Assuntos
Transportadores de Cassetes de Ligação de ATP/fisiologia , Apresentação de Antígeno , Antígenos de Histocompatibilidade Classe I/imunologia , Linfócitos T Citotóxicos/imunologia , Membro 2 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Substituição de Aminoácidos/genética , Animais , Anticorpos Bloqueadores/farmacologia , Antígenos CD8/imunologia , Antígenos de Histocompatibilidade Classe I/biossíntese , Células L , Camundongos , Camundongos Endogâmicos BALB C , Mutagênese Sítio-Dirigida , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Peptidil Dipeptidase A/fisiologia , Receptores de Antígenos de Linfócitos T/imunologia , Linfócitos T Citotóxicos/metabolismo , Vaccinia virus/imunologiaRESUMO
Genetic diversity and structure are described in the aquatic moss Platyhypnidium riparioides to assess its dispersal ability at a regional scale and to determine whether patterns of genetic differentiation correlate with environmental variation. Variation at six nuclear microsatellite loci from 50 populations in southern Belgium was investigated through Mantel tests, partial Mantel tests and spatial analysis of molecular variance. Overall patterns of genotypic variation showed strong differentiation among populations at a regional scale (F(ST) = 0.57). The high values of F(IS) observed within populations at both the ramet and genet levels, and the higher proportion of ramets with the same genotype than expected by chance, all point to a strongly clonal or selfing mating system. A genetic discontinuity was identified between northern and southern groups of populations. Within each group, F(ST) and geographical distances were significantly correlated. Partial Mantel tests suggest that genetic and ecological distances are significantly correlated in the southern group. The results point to strong dispersal limitation at the landscape scale and suggest that the southern and northern groups experienced different histories. Within the former, the correlation between genetic and ecological variation is suggestive of reproductive isolation among ecotypes.
Assuntos
Briófitas/genética , Fenômenos Ecológicos e Ambientais , Variação Genética , Água , Alelos , Bélgica , Frequência do Gene/genética , Geografia , Heterozigoto , Repetições de Microssatélites/genética , Ploidias , Dinâmica Populacional , Análise de Componente Principal , Tamanho da AmostraRESUMO
This paper documents the occurrence of allotriploidy (having three differentiated genomes) in gametophytes of two Southern Hemisphere Sphagnum species (S. australe, S. falcatulum). The pattern of microsatellite alleles indicates that both species are composed of a complex of allodiploid and allotriploid gametophytes, with the latter resulting from two allopolyploidization events. No haploid (n = x) gametophytes were found for either species. The ploidal levels suggested by the pattern of microsatellite alleles were confirmed by flow cytometry and Feulgen DNA image densitometry. For both S. australe and S. falcatulum, the respective allodiploid plants (or their ancestors) are one of the parent species of the allotriploid plants. This is the first report of triploidy in Sphagnum gametophytes occurring in nature and also the first report of the presence of three differentiated genomes in any bryophyte. It is also the first report of intersectional allopolyploidy in Sphagnum, with S. australe appearing to have parental species from Sphagnum sections Rigida and Sphagnum, and S. falcatulum having parental species from Sphagnum sections Cuspidata and Subsecunda. In both species, the allotriploid cytotypes were the most prevalent cytotype on the South Island of New Zealand. The pattern of microsatellite alleles shows the presence of two genetically distinct populations of allodiploid S. australe, possibly indicating multiple origins of polyploidy for that allodiploid cytotype. Morphological evidence is also highly indicative of recurrent polyploidy in the allotriploid cytotype of S. falcatulum. Allopolyploidy has clearly played a major evolutionary role in these two Southern Hemisphere taxa. This study, in conjunction with other recent research, indicates that allopolyploidy is a common, if not the predominant, form of polyploidy in Sphagnum.
Assuntos
Evolução Molecular , Genoma de Planta , Poliploidia , Sphagnopsida/genética , Alelos , DNA de Plantas/genética , Frequência do Gene , Genótipo , Repetições de Microssatélites , Nova Zelândia , Proteínas Proto-Oncogênicas c-met , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
Multiple paternity (polyandry) frequently occurs in flowering plants and animals and is assumed to have an important function in the evolution of reproductive traits. Polyandry in bryophytes may occur among multiple sporophytes of a female gametophyte; however, its occurrence and extent is unknown. In this study we investigate the occurrence and extent of multiple paternity, spatial genetic structure, and sporophytic inbreeding depression in natural populations of a dioicous bryophyte species, Sphagnum lescurii, using microsatellite markers. Multiple paternity is prevalent among sporophytes of a female gametophyte and male genotypes exhibit significant skew in paternity. Despite significant spatial genetic structure in the population, suggesting frequent inbreeding, the number of inbred and outbred sporophytes was balanced, resulting in an average fixation coefficient and population level selfing rate of zero. In line with the prediction of sporophytic inbreeding depression sporophyte size was significantly correlated with the level of heterozygosity. Furthermore, female gametophytes preferentially supported sporophytes with higher heterozygosity. These results indicate that polyandry provides the opportunity for postfertilization selection in bryophytes having short fertilization distances and spatially structured populations facilitating inbreeding. Preferential maternal support of the more heterozygous sporophytes suggests active inbreeding avoidance that may have significant implications for mating system evolution in bryophytes.
Assuntos
Briófitas/genética , Briófitas/fisiologia , Fertilização/fisiologia , Endogamia , Pólen/fisiologia , Fertilização/genética , Genes de Plantas , Variação Genética/fisiologia , Genética Populacional , Óvulo Vegetal/genética , Óvulo Vegetal/fisiologia , Pólen/genética , Especificidade da EspécieRESUMO
Several lines of evidence suggest that recent long-distance dispersal may have been important in the evolution of intercontinental distribution ranges of bryophytes. However, the absolute rate of intercontinental migration and its relative role in the development of certain distribution ranges is still poorly understood. To this end, the genetic structure of intercontinental populations of six peatmoss species showing an amphi-Atlantic distribution was investigated using microsatellite markers. Methods relying on the coalescent were applied (IM and MIGRATE) to understand the evolution of this distribution pattern in peatmosses. Intercontinental populations of the six peatmoss species were weakly albeit significantly differentiated (average F(ST) = 0.104). This suggests that the North Atlantic Ocean is acting as a barrier to gene flow even in bryophytes adapted to long-range dispersal. The im analysis suggested a relatively recent split of intercontinental populations dating back to the last two glacial periods (9000-289,000 years ago). In contrast to previous hypotheses, analyses indicated that both ongoing migration and ancestral polymorphism are important in explaining the intercontinental genetic similarity of peatmoss populations, but their relative contribution varies with species. Migration rates were significantly asymmetric towards America suggesting differential extinction of genotypes on the two continents or invasion of the American continent by European lineages. These results indicate that low genetic divergence of amphi-Atlantic populations is a general pattern across numerous flowering plants and bryophytes. However, in bryophytes, ongoing intercontinental gene flow and retained shared ancestral polymorphism must both be considered to explain the genetic similarity of intercontinental populations.
Assuntos
Genética Populacional , Filogenia , Polimorfismo Genético , Sphagnopsida/genética , Alelos , Europa (Continente) , Evolução Molecular , Fluxo Gênico , Marcadores Genéticos , Especiação Genética , Repetições de Microssatélites , Modelos Genéticos , Mutação , América do Norte , Sphagnopsida/classificaçãoRESUMO
Allopolyploidy is probably the most extensively studied mode of plant speciation and allopolyploid species appear to be common in the mosses (Bryophyta). The Sphagnum subsecundum complex includes species known to be gametophytically haploid or diploid, and it has been proposed that the diploids (i.e., with tetraploid sporophytes) are allopolyploids. Nucleotide sequence and microsatellite variation among haploids and diploids from Newfoundland and Scandinavia indicate that (1) the diploids exhibit fixed or nearly fixed heterozygosity at the majority of loci sampled, and are clearly allopolyploids, (2) diploids originated independently in North America and Europe, (3) the European diploids appear to have the haploid species, S. subsecundum, as the maternal parent based on shared chloroplast DNA haplotypes, (4) the North American diploids do not have the chloroplast DNA of any sampled haploid, (5) both North American and European diploids share nucleotide and microsatellite similarities with S. subsecundum, (6) the diploids harbor more nucleotide and microsatellite diversity than the haploids, and (7) diploids exhibit higher levels of linkage disequilibrium among microsatellite loci. An experiment demonstrates significant artifactual recombination between interspecific DNAs coamplified by PCR, which may be a complicating factor in the interpretation of sequence-based analyses of allopolyploids.
Assuntos
Evolução Molecular , Especiação Genética , Sphagnopsida/classificação , Sphagnopsida/genética , Teorema de Bayes , DNA de Cloroplastos/genética , DNA de Plantas/genética , Diploide , Europa (Continente) , Genética Populacional , Haploidia , Haplótipos , Heterozigoto , Funções Verossimilhança , Repetições de Microssatélites , América do Norte , Filogenia , Polimorfismo Genético , Poliploidia , Análise de Componente Principal , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Quantitative genetic analyses of Drosophila melanogaster stocks with postponed aging have suffered from the problem of a lack of certainty concerning patterns of allelic differentiation. The present experiments were designed to alleviate this difficulty by selecting for enhanced levels of characters known to be related to postponed aging. Selection successfully increased the degree of differentiation of postponed aging stocks with respect to starvation resistance and fecundity, but persistent additive genetic variance suggested that selection did not result in fixation of alleles. The artificially selected stocks were subjected to crosses to test for patterns of dominance and maternal effects. There was little evidence for these effects in the inheritance of the characters underlying postponed aging, even with the increased differentiation of the selected stocks.
Assuntos
Drosophila melanogaster/genética , Variação Genética , Envelhecimento/genética , Alelos , Análise de Variância , Animais , Cruzamentos Genéticos , Drosophila melanogaster/fisiologia , Feminino , Fertilidade/genética , Genes Dominantes/genética , Longevidade/genética , Masculino , Polimorfismo Genético , Seleção Genética , InaniçãoRESUMO
Pleurocarpous mosses, characterized by lateral female gametangia and highly branched, interwoven stems, comprise three orders and some 5000 species, or almost half of all moss diversity. Recent phylogenetic analyses resolve the Ptychomniales as sister to the Hypnales plus Hookeriales. Species richness is highly asymmetric with approximately 100 Ptychomniales, 750 Hookeriales, and 4400 Hypnales. Chloroplast DNA (cpDNA) sequences were obtained to compare partitioning of molecular diversity among the orders with estimates of species richness, and to test the hypothesis that either the Hookeriales or Hypnales underwent a period (or periods) of exceptionally rapid diversification. Levels of biodiversity were quantified using explicitly historical "phylogenetic diversity" and non-historical estimates of standing sequence diversity. Diversification rates were visualized using lineage-through-time (LTT) plots, and statistical tests of alternative diversification models were performed using the methods of Paradis (1997). The effects of incomplete sampling on the shape of LTT plots and performance of statistical tests were investigated using simulated phylogenies with incomplete sampling. Despite a much larger number of accepted species, the Hypnales contain lower levels of (cpDNA) biodiversity than their sister group, the Hookeriales, based on all molecular measures. Simulations confirm previous results that incomplete sampling yields diversification patterns that appear to reflect a decreasing rate through time, even when the true phylogenies were simulated with constant rates. Comparisons between simulated results and empirical data indicate that a constant rate of diversification cannot be rejected for the Hookeriales. The Hypnales, however, appear to have undergone a period of exceptionally rapid diversification for the earliest 20% of their history.
Assuntos
Biodiversidade , Briófitas/genética , Evolução Molecular , Modelos Genéticos , Filogenia , Sequência de Bases , DNA de Cloroplastos/genética , Funções Verossimilhança , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
Mechanisms of initial hypercalcaemic responses to parathyroid hormone (PTH) and 16,16-dimethyl prostaglandin (PG) E2 have been investigated in 10-to 12-day-old chicks in vivo using a combination of acute 45Ca injection and microwave fixation to stabilize tissue isotope levels. Single i.v. injection of 16,16-dimethyl PGE2 (20 micrograms/100 g body wt) caused an approximately 100% increase in soft tissue 45Ca levels compared with vehicle control injected chicks at 30 min. 45Ca levels were lowered in calvarium by 26% and in femur by 60% with this treatment. Bovine PTH (1-34) (3.3 micrograms/100 g body wt) had no effect on soft tissue 45Ca levels, but in calvarium it had a similar effect to the PG. In femur this dose of PTH lowered 45Ca by 19%. When expressed on an absolute basis (c.p.m./100 mg tissue wt), responses to the PG in soft tissue were only 3 and 10% respectively of those in femur and calvarium. The duration of inhibitory responses in bone were examined and those to PTH found to be transient (less than 45 min) compared with the responses to the PG (greater than 135 min). Dose-response curves for PTH- and PG-induced inhibition of 45Ca uptake into femur at 15 min were essentially parallel and indicated that the lowest doses of PTH and PG used (0.74 micrograms and 1.1 micrograms/100 body wt respectively) produced significant responses. In a separate experiment it was found that inhibition of 45Ca uptake into femur was evident as early as 3 min following PTH or PG injection.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
16,16-Dimetilprostaglandina E2/farmacologia , Osso e Ossos/metabolismo , Cálcio/metabolismo , Hormônio Paratireóideo/farmacologia , Fragmentos de Peptídeos/farmacologia , Prostaglandinas E Sintéticas/farmacologia , Animais , Osso e Ossos/efeitos dos fármacos , Galinhas , Relação Dose-Resposta a Droga , Fêmur/metabolismo , Fígado/metabolismoRESUMO
The rapid effects of parathyroid hormones and a variety of prostaglandins on net uptake of 45Ca into the skeleton have been investigated in chicks and, in a limited parallel study, in immature rats. Intravenous injection of bovine (b) parathyroid hormone(1-34) (bPTH(1-34)) or 16,16-dimethyl prostaglandin E2 (16,16-dimethyl PGE2) in a 45Ca-labelled vehicle, combined with subsequent microwave fixation of tissue isotope levels, resulted in rapid (3-15 min) net inhibition of 45Ca uptake into endochondral bone (femur) in chicks (12 days old) and rats (4 weeks old). Use of 125I-labelled albumin and [14C]mannitol indicated that these responses were not a reflection of gross changes in tissue vascular or extracellular space. In rats, bPTH(1-84) also caused significant net inhibition of 45Ca uptake into femur at 10 min. Both bPTH(1-34) and 16,16-dimethyl PGE2 produced generally smaller decreases in 45Ca uptake into chick dermal bone (calvarium) at 3-15 min. In rat calvarium, however, these agents stimulated net uptake of 45Ca at these times. When microwave fixation was omitted, inhibitory responses were reduced or disappeared, while the stimulatory response in rat calvarium was enhanced. Responses to natural prostaglandins (PGE1, PGE2, PGF2 alpha and PGI2) in chicks at 3 min were similar but less marked than those to 16,16-dimethyl PGE2; 45Ca uptake into femur and, to a lesser extent in calvarium, being inhibited. In rats, PGE1, PGE2 and PGF2 alpha showed a tendency to decrease 45Ca uptake into femur while PGE1 and PGE2 both increased 45Ca uptake into calvarium.
Assuntos
Osso e Ossos/metabolismo , Cálcio/metabolismo , Hormônios/farmacologia , Hormônio Paratireóideo/farmacologia , Fragmentos de Peptídeos/farmacologia , Prostaglandinas E Sintéticas/farmacologia , Animais , Osso e Ossos/efeitos dos fármacos , Galinhas , Feminino , Masculino , RatosRESUMO
Two human parathyroid hormone-related protein (hPTHrP) fragments were tested for effects on maternofetal transfer of 45Ca and Mg across the in-situ perfused rat placenta at 21 days of gestation (term = 23 days). The fetal placental circulation was perfused with a Mg-free Krebs-Ringer solution and the unidirectional maternofetal clearance (Kmf) of 45Ca and Mg compared with that of 51Cr-EDTA, the latter being employed as a paracellular diffusional marker. Placental perfusion with hPTHrP(1-34) (100 ng/ml) or hPTHrP(75-86)amide (50 ng/ml) did not significantly alter the Kmf of 45Ca or that of Mg. In separate rats, however, hPTHrP(1-34) but not hPTHrP(75-86)amide stimulated marked placental cyclic AMP (cAMP) release, the peak response of 63 +/- 7 pmol/min occurring 10 min after the beginning of the peptide perfusion. A lower dose of hPTHrP(1-34) (4 ng/ml) produced a similar peak release of cAMP, as did [Nle8,21, Tyr34]-rPTH(1-34)amide (4 ng/ml) and the adenylate cyclase agonist forskolin (17 mumol/l). Forskolin also rapidly increased the Kmf of 45Ca but not that of Mg or 51Cr-EDTA. The present study indicates that hPTHrP does not acutely affect maternofetal transfer of Ca or Mg across the perfused rat placenta. The data also question the role played by cAMP in the stimulatory actions of forskolin on placental Ca transport.
Assuntos
Cálcio/metabolismo , AMP Cíclico/biossíntese , Magnésio/metabolismo , Troca Materno-Fetal/efeitos dos fármacos , Proteína Relacionada ao Hormônio Paratireóideo , Hormônio Paratireóideo/farmacologia , Fragmentos de Peptídeos/farmacologia , Placenta/metabolismo , Animais , Colforsina/farmacologia , Feminino , Técnicas de Cultura de Órgãos , Perfusão , Placenta/efeitos dos fármacos , Gravidez , Proteínas da Gravidez/farmacologia , Proteínas/farmacologia , Ratos , Ratos EndogâmicosRESUMO
Two techniques for the automatic selection of individual leaf positions of a Philips multi-leaf collimator are described. Target volumes are identified either on simulator images or on cross-sectional images from CT or MR scanners. The setting of each leaf is computed to position the beam edge to cover the target with an appropriate, user defined, margin. An important consideration in the development of the system was its robustness and so the applications initially implemented have been relatively simple, comprising single field, parallel opposed fields and coplanar 4 field box techniques. Attention has been paid to the overall integrity of the planning and treatment delivery process. Before treatment commences, the beam shapes, which have been generated by the computer and transferred to the MLC control computer over a local area network, are checked against a printed template representing each beam. All data used for planning is archived and is accessible for review or, if necessary, for treatment modification.
Assuntos
Planejamento da Radioterapia Assistida por Computador/métodos , Humanos , Imageamento por Ressonância Magnética , Tomografia Computadorizada por Raios XRESUMO
Two-year-old red spruce (Picea rubens Sarg.), raised from seed collected at five locations from Maine to North Carolina, were exposed in a greenhouse to either sulphuric or nitric acid mist (median pH 2-85). Exposures were performed three times a day, four days a week over an eleven-week period during the growing season. Each exposure consisted of 4 h of misting followed by a 3.75 h drying period. Immediately before each drying period, acidic droplets were either left to dry directly on the foliage or were removed by a 15 min deionized water (diw) rinse. When sulphuric acid droplets dried directly on the foliage, visible injury developed and to a lesser degree the same was true for nitric acid. When acidic droplets were replaced with dim before each drying period, foliar injury did not develop despite misting for hundreds of hours at pH values well within the range that has caused visible injury in previous experiments. When acidic droplets dried directly on foliage, needle dry weight was reduced, but differences between sulphuric and nitric acid exposure were not found. No treatment effects were found on terminal shoot length. Treatment responses among seedlings from different seed resources were similar, with greater variation found within provenances than between provenances. We conclude that the composition of liquid on foliage at the end of wet deposition events and the processes occurring during drying periods are important factors in determining the response of red spruce to acidic cloudwater.
RESUMO
Bryophytes, including the mosses, liverworts, and hornworts, occur in a variety of habitats with high concentrations of metals and have other characteristics that are advantageous for studies of metal tolerance. Mosses may evolve genetically specialized, metal-tolerant races less frequently than flowering plants. Some species of mosses appear to have inherently high levels of metal tolerance even in individuals that have not been subjected to natural selection in contaminated environments. Scopelophila cataractae, one of the so-called copper mosses, not only tolerates extremely high concentrations of metals in its substrates, but requires these substrates for optimum growth. This species should be included in mechanistic studies of tolerance at the cellular and molecular levels.